PROJECT SUMMARY: Metabolomics Shared Resource (MSR) The Metabolomics Shared Resource (MSR) is evolving to fulfill the research needs and mission of the Dan L Duncan Comprehensive Cancer Center (DLDCCC). This Shared Resource (SR) provides advanced mass spectrometry approaches and technologies, scientific consultation, and expert data analytics as is required for high quality global and targeted metabolomics studies. The MSR specializes in discovery, identification, characterization, and quantification of biomolecules, as well as metabolic flux analysis, from a variety of biological specimens including tissues, cell lines, and fluids. The main infrastructure of the MSR consists of 5 high-end mass spectrometry systems and upfront liquid chromatography columns that are most suitable for covering the range of metabolomic analyses developed as services including an Agilent 6490 Triple Quad Mass Spectrometer, 2 Agilent 6495 Triple Quad Mass Spectrometers, an Agilent 6550 Time of Flight Mass Spectrometer, and a SCIEX Triple TOF mass spectrometer with Schimadzu Nexera UHPLC-System with data- independent acquisition for lipidomics. The MSR has a dedicated team of experts in mass spectrometry with extensive experience in metabolomic profiling and biological expertise in cellular metabolism and data analysis. Metabolomics became a Shared Resource of the Cancer Center for the first time at the last competing renewal of the CCSG in 2015. Since then, the MSR has supported a broad range of DLDCCC members across 6 of the 7 Programs and a total of 57 publications, many representing major projects in high- impact journals including Nature, Journal of Clinical Investigations, Nature Communications, PLoS Genetics, Nature Metabolism, Cell Reports, Cancer Cell, Cancer Research, and Clinical Cancer Research. This is a dynamic SR facility that has kept pace with rapidly emerging technologies and instrumentation and has been a leader in the field by developing and publishing innovative technologies. The MSR has developed state-of-the- art technologies as core support services that includes the following: (1) Steady-state targeted metabolomics with a capacity for quantification of up to 700 known metabolites; (2) Untargeted metabolomics for identification of up to 1200 metabolites; (3) Lipidomics for up to 800 molecules and; (4) Metabolic flux analysis to monitor changes in activities of major metabolic pathways in experimental model systems. The MSR, as proposed in this application, can bridge the gap in our understanding of molecular pathways involved in cancer biology and enable a wide range of DLDCCC research groups to explore the role of metabolism in cancer utilizing cutting- edge technologies with quality, efficiency, and accuracy which would not be possible otherwise.